Impact of bacterial admixtures on the compressive and tensile strengths, permeability, and pore structure of ternary mortars: Comparative study of ureolytic and phototrophic bacteria.

The addition of bacterial biomass to cementitious materials can improve strength and permeability properties by altering the pore structure. Photoautotrophic bacteria are understudied mortar bio-additives that do not produce unwanted by-products compared to commonly studied ureolytic species. This study directly compares the impact of the addition of heterotrophic Bacillus subtilis to photoautotrophic Synechocystis sp. PCC6803 on mortar properties and microstructure. Cellulose fibers were used as a bacteria carrier. A commercial concrete healing agent composed of dormant bacterial spores was also tested. Strength, water absorption tests, mercury intrusion porosimetry, differential scanning calorimetry, thermogravimetric analysis, and scanning electron microscopy were applied to experimental mortar properties. The photoautotrophic modifications had a stronger positive impact on mortar strength and permeability properties than sporulated heterotrophic modifications due to differences in surface properties and production of exopolysaccharides. The findings provide support for photoautotrophic species as additives for mortars to move away from ammonia-generating species.

Exploring a high-urease activity Bacillus cereus for self-healing concrete via induced CaCO3 precipitation.

The structural integrity and esthetic appeal of concrete can be compromised by concrete cracks. Promise has been shown by microbe-induced calcium carbonate precipitation (MICP) as a solution for concrete cracking, with a focus on urease-producing microorganisms in research. Bacillus cereus was isolated from soil and employed for this purpose in this study due to its high urease activity. The strain exhibited strong tolerance for alkaline media and high salt levels, which grew at a pH of 13 and 4% salt concentration. The repair of concrete cracks with this strain was evaluated by assessing the effects of four different thickeners at varying concentrations. The most effective results were achieved with 10 g/L of sodium carboxymethyl cellulose (CMC-Na). The data showed that over 90% repair of cracks was achieved by this system with an initial water penetration time of 30 s. The study also assessed the quantity and sizes of crystals generated during the bacterial mineralization process over time to improve our understanding of the process. KEY POINTS: • MICP using Bacillus cereus shows potential for repairing concrete cracks. • Strain tolerates alkaline media and high salt levels, growing at pH 13 and 4% salt concentration. • Sodium carboxymethyl cellulose (CMC-Na) at 10 g/L achieved over 90% repair of cracks.

Effect of Bacillus subtilis on mechanical and self-healing properties in mortar with different crack widths and curing conditions.

This research aimed to investigate the effectiveness of Bacillus subtilis (B. subtilis) in self-healing cracks in concrete and enhancing concrete strength through microbial induced calcium carbonate precipitation (MICP). The study evaluated the ability of the mortar to cover cracks within 28 days, taking into account the width of the crack, and observed the recovery of strength after self-healing. The use of microencapsulated endospores of B. subtilis was also examined for its impact on the strength of concrete. The compressive, splitting tensile, and flexural strengths of normal mortar were compared to those of biological mortar, and it was found that biological mortar had a higher strength capacity. Microstructure analysis using SEM and EDS showed that bacterial growth increased calcium production, contributing to the improved mechanical properties of the bio-mortar.

Use of bacterial binder in repair mortar for micro-crack remediation.

Micro-cracks are one of the types of stone deterioration which can propagate and lead to surface detachments and larger cracks in the long run. The present study developed a sustainable and environmentally friendly infill material-biological mortar (BM), as an alternative to conventional approaches. Using a biomineralization approach, this BM was explicitly designed for healing micro-cracks (less than 2 mm) in historic travertines. To this end, the mortar was prepared using a calcifying Bacillus sp. isolated from thermal spring water resources in Pamukkale Travertines (Denizli), stone powder gathered from travertine quarries in the vicinity, and a triggering solution specifically designed to set off calcium carbonate precipitation reaction. After setup, BM was applied to micro-cracks of artificially aged test stones for testing. Scanning electron microscopy revealed calcium carbonate-coated Bacillus sp. bodies in the BM matrix, optical microscopy showed secondary calcite minerals throughout the BM applied micro-cracks, and stereomicroscopy and nanoindentation analyses demonstrated bonding of BM with stone due to microbial calcification activities. Furthermore, BM and original material contact showed a continuous and coherent structure in all samples. Within this context, BM could be considered a promising and alternative approach for the remediation of micro-cracks of historic stones. KEY POINTS: A binder was produced by the MICP of Bacillus sp. Pamukkale. Physical, mineralogical, and nanomechanical characterization demonstrated microbial calcite precipitates in BM. A significant bond was determined between the grains and matrix of BM due to Bacillus sp. calcite production activities.

Impact of the immobilized Bacillus cereus MG708176 on the characteristics of the bio-based self-healing concrete.

Novel carrier units were evaluated for their bio-healing benefits in our study to increase the efficacy of concrete healing. Bacillus cereus MG708176, an alkali-tolerant, calcite precipitating, endospore-forming strain was added as a bio-healing agent after its immobilization on wood ash units. A spore concentration of [1.3 × 107 spore/cm3] combined with 2.5% w/w urea was added to cement. Beams of 40 × 40 × 160 mm were used and tested for completely damaged mortar specimens after 7, 14, and 28 days of water treatment. Using wood ash bacterial mortars, totally destructed specimens were fully healed in all time intervals. Positive changes in concrete mechanical properties in bacterial wood ash treatment that were 24.7, 18.9, and 28.6% force for compressive, flexural, and tensile strengths more than control. The micro-images of the Scanning Electron Microscope (SEM) showed the dense concrete structure via calcite, Bacillafilla, and ettringite formation. Our results have shown improvements in the concrete healing efficiency and the mechanical concrete properties by filling the concrete cracks using a calcite-producing bacterium that is immobilized on wood ash units.

Performance of biological methods on self-healing and mechanical properties of concrete using S. pasteurii.

Biological methods (adding bacteria to the concrete mixtures) among the most recently investigated procedures increase the durability of concrete and repair concrete cracks. In the present study, different biological methods were used to heal the cracks of concrete and the most suitable method was subsequently introduced as the main aim of the research. For this purpose, the culture medium, various sources of calcium salts as bacterial nutrients, and the effect of air-entrained agent on the healing process were studied. The results showed that the use of bacterial nutrient inside the concrete mixes has an affirmative impact on the mechanical properties and self-healing characteristics of concretes. Simultaneous use of Sporosarcina pasteurii bacteria and calcium nitrate-urea or calcium chloride-urea as a bacterial nutrient in the concrete mixture increased the 28 days compressive strength of concretes by 23.4% and 7.5%, respectively. The utilization of bacterial cells, nutrients, and culture in the concrete mixture provided the ability to heal wide cracks where the healing time was significantly reduced (about 8 days). On the other hand, separation of the bacterial culture medium slightly reduced the self-healing performance of the concretes.

Performance Evaluation of Bio Concrete by Cluster and Regression Analysis for Environment Protection.

The focus of this research is to isolating and identifying bacteria that produce calcite precipitate, as well as determining whether or not these bacteria are suitable for incorporation into concrete in order to enhance the material's strength and make the environment protection better. In order to survive the high "potential of hydrogen" of concrete, microbes that are going to be added to concrete need to be able to withstand alkali, and they also need to be able to develop endospores so that they can survive the mechanical forces that are going to be put on the concrete while it is being mixed. In order to precipitate CaCO3 in the form of calcite, they need to have a strong urease activity. Both Bacillus sphaericus and the Streptococcus aureus bacterial strains were evaluated for their ability to precipitate calcium carbonate (CaCO3). These strains were obtained from the Department of Biotechnology at GLA University in Mathura. This research aims to solve the issue of augmenting the tension and compression strengths of concrete by investigating possible solutions for environmentally friendly concrete. The sterile cultures of the microorganisms were mixed with water, which was one of the components of the concrete mixture, along with the nutrients in the appropriate proportions. After that, the blocks were molded, and then pond-cured for 7, 28, 56, 90, 120, 180, 270, and 365 days, respectively, before being evaluated for compressibility and tensile strength. An investigation into the effect that bacteria have on compression strength was carried out, and the outcomes of the tests showed that bacterial concrete specimens exhibited an increase in mechanical strength. When compared to regular concrete, the results showed a maximum increase of 16 percent in compressive strength and a maximum increase of 12 percent in split tensile strength. This study also found that both bacterial concrete containing 106, 107, and 108 cfu/ml concentrations made from Bacillus sphaericus and Streptococcus aureus bacteria gave better results than normal concrete. Both cluster analysis (CA) and regression analysis (RA) were utilized in this research project in order to measure and analyze mechanical strength.

Strain Screening and Particle Formation: a Lysinibacillus boronitolerans for Self-Healing Concrete.

Microbial-induced calcite precipitation is a promising technology to solve the problem of cracks in soil concrete. The most intensively investigated microorganisms are urease-producing bacteria. Lysinibacillus that is used as urease-producing bacteria in concrete repair has rarely been reported. In this study, Lysinibacillus boronitolerans with a high urease activity was isolated from soil samples. This strain is salt- and alkali-tolerance, and at pH 13, can grow to ~OD600 2.0 after 24 h. At a salt concentration of 6%, the strain can still grow to ~OD600 1.0 after 24 h. The feasibility of using this strain in self-healing concrete was explored. The data showed that cracks within ~0.6 mm could be repaired naturally with hydration when spores and substrates were added to the concrete in an appropriate proportion. Moreover, the number and morphology of CaCO3 crystals that were produced by bacteria can be influenced by the concrete environment. An efficiency method to elucidate the process of microbial-induced calcium carbonate crystal formation was established with Particle Track G400. This study provides a template for future studies on the theory of mineralization based on microorganisms. IMPORTANCE The formation of calcium carbonate crystals in concrete by urease-producing bacteria is not understood fully. In this study, a Lysinibacillus boronitolerans strain with a high urease activity was isolated and used to analyze the counts and sizes of the crystals and the relationship with time. The data showed that the number of crystal particles increases exponentially in a short period with sufficient substrate, after which the crystals grow, precipitate or break. In concrete, the rate-limiting steps of calcium carbonate crystal accumulation are spore germination and urease production. These results provided data support for the rational design of urease-producing bacteria in concrete repair.

Analysis of mold and mycotoxins in naturally infested indoor building materials.

Health issues of residents of mold-infested housing are reported on a regular basis, and reasons for the arising impairments can be manifold. One possible cause are the toxic secondary metabolite produced by indoor microfungi (mycotoxins). To enable a more thorough characterization of the exposure to mycotoxins in indoor environments, data on occurrence and quantities of mycotoxins is essential. In the presented study, 51 naturally mold-infested building material samples were analyzed applying a previously developed method based on ultra-high performance liquid chromatography (UHPLC) separation in combination with triple-quadrupole mass spectrometry (TQMS) detection. A total of 38 secondary metabolites derived from different indoor mold genera like Aspergillus, Fusarium, Penicillium, and Stachybotrys were analyzed, of which 16 were detectable in 28 samples. As both the spectrum of target analytes and the investigated sample matrices showed high chemical varieties, an alternative calibration approach was applied complementary to identify potentially emerging matrix effects during ionization and mass spectrometric detection. Overall, strong alterations of analyte signals were rare, and compensation of considerable matrix suppression/enhancement only had to be performed for certain samples. Besides mycotoxin determination and quantification, the presence of 18 different mold species was confirmed applying microbiological approaches in combination with macro- and microscopic identification according to DIN ISO 16000-17:2010-06. These results additionally highlight the diversity of mycotoxins potentially arising in indoor environments and leads to the assumption that indoor mycotoxin exposure stays an emerging topic of research, which has only just commenced.

Physiological suitability of sulfate-reducing granules for the development of bioconcrete.

Regular monitoring and timely repair of concrete cracks are required to minimize further deterioration. Self-healing of cracks has been proposed as an alternative to the crack maintenance procedures. One of the proposed techniques is to use axenic cultures to exploit microbial-induced calcite precipitation (MICP). However, such healing agents are not cost-effective for in situ use. As the market for bio-based self-healing concrete necessitates a low-cost bio-agent, nonaxenic sulfate reducing bacterial (SRB) granules were investigated in this study through cultivation in an upflow anaerobic sludge blanket reactor. The compact granules can protect the bacteria from adverse conditions without encapsulation. This study investigated the microbial activities of SRB granules at different temperatures, pH, and chemical oxygen demand concentrations which the microbes would experience during the concrete casting and curing process. The attenuation and recovery of microbial activities were measured before and after the exposure. Moreover, the MICP yield was also tested for a possible use in self-healing bioconcrete. The results consistently showed that SRB granules were able to survive starvation, high temperature (50-60°C), and high pH (12), together with scanning electron microscope/energy dispersive spectrometry/X-ray diffraction analysis evidence. Microbial staining analysis demonstrated the formation of spores in the granules during their exposure to harsh conditions. SRB granule was thus demonstrated to be a viable self-healing nonaxenic agent for low-cost bioconcrete.

A critical review on microbial carbonate precipitation via denitrification process in building materials.

The naturally occurring biomineralization or microbially induced calcium carbonate (MICP) precipitation is gaining huge attention due to its widespread application in various fields of engineering. Microbial denitrification is one of the feasible metabolic pathways, in which the denitrifying microbes lead to precipitation of carbonate biomineral by their basic enzymatic and metabolic activities. This review article explains all the metabolic pathways and their mechanism involved in the MICP process in detail along with the benefits of using denitrification over other pathways during MICP implementation. The potential application of denitrification in building materials pertaining to soil reinforcement, bioconcrete, restoration of heritage structures and mitigating the soil pollution has been reviewed by addressing the finding and limitation of MICP treatment. This manuscript further sheds light on the challenges faced during upscaling, real field implementation and the need for future research in this path. The review concludes that although MICP via denitrification is an promising technique to employ it in building materials, a vast interdisciplinary research is still needed for the successful commercialization of this technique.

Attachment on mortar surfaces by cyanobacterium Gloeocapsa PCC 73106 and sequestration of CO2 by microbially induced calcium carbonate.

Cyanobacterial carbonate precipitation induced by cells and extracellular polymeric substances (EPS) enhances mortar durability. The percentage of cell/EPS attachment regulates the effectiveness of the mortar restoration. This study investigates the cell coverage on mortar and microbially induced carbonate precipitation. Statistical analysis of results from scanning electron and fluorescence microscopy shows that the cell coverage was higher in the presence of UV-killed cells than living cells. Cells are preferably attached to cement paste than sand grains, with a difference of one order of magnitude. The energy-dispersive X-ray spectroscopy analyses and Raman mapping suggest cyanobacteria used atmospheric CO2 to precipitate carbonates.

Extremophilic taxa predominate in a microbial community of photovoltaic panels in a tropical region.

Photovoltaic panels can be colonized by a highly diverse microbial diversity, despite life-threatening conditions. Although they are distributed worldwide, the microorganisms living on their surfaces have never been profiled in tropical regions using 16S rRNA high-throughput sequencing and PICRUst metagenome prediction of functional content. In this work, we investigated photovoltaic panels from two cities in southeast Brazil, Sorocaba and Itatiba, using these bioinformatics approach. Results showed that, despite significant differences in microbial diversity (p < 0.001), the taxonomic profile was very similar for both photovoltaic panels, dominated mainly by Proteobacteria, Bacteroidota and lower amounts of Cyanobacteria phyla. A predominance of Hymenobacter and Methylobacterium-Methylorubrum was observed at the genus level. We identified a microbial common core composed of Hymenobacter, Deinococcus, Sphingomonas, Methylobacterium-Methylorubrum, Craurococcus-Caldovatus, Massilia, Noviherbaspirillum and 1174-901-12 sharing genera. Predicted metabolisms focused on specific genes associated to radiation and desiccation resistance and pigments, were detected in members of the common core and among the most abundant genera. Our results suggested that taxonomic and functional profiles investigated were consistent with the harsh environment that photovoltaic panels represent. Moreover, the presence of stress genes in the predicted functional content was a preliminary evidence that microbes living there are a possibly source of metabolites with biotechnological interest.

Improvement of Biomineralization of Sporosarcina pasteurii as Biocementing Material for Concrete Repair by Atmospheric and Room Temperature Plasma Mutagenesis and Response Surface Methodology.

Microbially induced calcium carbonate precipitation (MICP) has recently become an intelligent and environmentally friendly method for repairing cracks in concrete. To improve on this ability of microbial materials concrete repair, we applied random mutagenesis and optimization of mineralization conditions to improve the quantity and crystal form of microbially precipitated calcium carbonate. Sporosarcina pasteurii ATCC 11859 was used as the starting strain to obtain the mutant with high urease activity by atmospheric and room temperature plasma (ARTP) mutagenesis. Next, we investigated the optimal biomineralization conditions and precipitation crystal form using Plackett-Burman experimental design and response surface methodology (RSM). Biomineralization with 0.73 mol/l calcium chloride, 45 g/l urea, reaction temperature of 45°C, and reaction time of 22 h, significantly increased the amount of precipitated calcium carbonate, which was deposited in the form of calcite crystals. Finally, the repair of concrete using the optimized biomineralization process was evaluated. A comparison of water absorption and adhesion of concrete specimens before and after repairs showed that concrete cracks and surface defects could be efficiently repaired. This study provides a new method to engineer biocementing material for concrete repair.

Characterization of a Novel CaCO3-Forming Alkali-Tolerant Rhodococcus erythreus S26 as a Filling Agent for Repairing Concrete Cracks.

Biomineralization, a well-known natural phenomenon associated with various microbial species, is being studied to protect and strengthen building materials such as concrete. We characterized Rhodococcus erythreus S26, a novel urease-producing bacterium exhibiting CaCO3-forming activity, and investigated its ability in repairing concrete cracks for the development of environment-friendly sealants. Strain S26 grown in solid medium formed spherical and polygonal CaCO3 crystals. The S26 cells grown in a urea-containing liquid medium caused culture fluid alkalinization and increased CaCO3 levels, indicating that ureolysis was responsible for CaCO3 formation. Urease activity and CaCO3 formation increased with incubation time, reaching a maximum of 2054 U/min/mL and 3.83 g/L, respectively, at day four. The maximum CaCO3 formation was achieved when calcium lactate was used as the calcium source, followed by calcium gluconate. Although cell growth was observed after the induction period at pH 10.5, strain S26 could grow at a wide range of pH 4-10.5, showing its high alkali tolerance. FESEM showed rhombohedral crystals of 20-60 µm in size. EDX analysis indicated the presence of calcium, carbon, and oxygen in the crystals. XRD confirmed these crystals as CaCO3 containing calcite and vaterite. Furthermore, R. erythreus S26 successfully repaired the artificially induced large cracks of 0.4-0.6 mm width.

The theoretical adhesion of Pseudomonas aeruginosa and Escherichia coli on some plumbing materials in presence of distilled water or tap water.

The main aim of this work was to determine the most appropriate materials for the installation of a water system according to the characteristics of the water that passes through it. To this end, we conducted an investigation of the effect of two types of water (SDW: sterile distilled water and STW: sterile tap water) on the properties of bacterial surfaces and the theoretical adhesion of two bacteria (Pseudomonas aeruginosa and Escherichia coli) on six plumbing materials. Contact angle measurements were used to determine the surface energies of bacteria and materials. XDLVO theory was used to estimate the interactions between bacteria and plumbing materials. The results showed that water had a clear impact on the electron donor character and the hydrophobicity of the bacterial surfaces. Also, the predictive adhesion showed that all tested materials could be colonized by P. aeruginosa and E. coli ([Formula: see text]<0). However, colonization became thermodynamically less favorable or unfavorable (increase in [Formula: see text] values) with SDW and STW, respectively. Finally, the results suggest that the choice of the most suitable material for a drinking water installation is related to the quality of the water itself.

Calcite seed-assisted microbial induced carbonate precipitation (MICP).

Microbial-induced calcium carbonate precipitation (MICP) is a biological process inducing biomineralization of CaCO3. This can be used to form a solid, concrete-like material. To be able to use MICP successfully to produce solid materials, it is important to understand the formation process of the material in detail. It is well known that crystallization surfaces can influence the precipitation process. Therefore, we present in this contribution a systematic study investigating the influence of calcite seeds on the MICP process. We focus on the changes in the pH and changes of the optical density (OD) signal measured with absorption spectroscopy to analyze the precipitation process. Furthermore, optical microscopy was used to visualize the precipitation processes in the sample and connect them to changes in the pH and OD. We show, that there is a significant difference in the pH evolution between samples with and without calcite seeds present and that the shape of the pH evolution and the changes in OD can give detailed information about the mineral precipitation and transformations. In the presented experiments we show, that amorphous calcium carbonate (ACC) can also precipitate in the presence of initial calcite seeds and this can have implications for consolidated MICP materials.

Microbial effect on water sorptivity and sulphate ingress by Bacillus megaterium on mortars prepared using Portland Pozzolana cement.

AIMS: To determine the effect of direct embedment of Bacillus megaterium into Portland pozzolana cement mortars on water sorptivity and diffusivity coefficient of sulphate ions. METHODS AND RESULTS: Prisms with a water/cement ratio of 0·5 were prepared by blending Portland Pozzolana cement with the requisite volume of a B. megaterium (microbial) solution whose concentration was 1·0 × 107 cells per ml. Mortar prisms of 160 mm × 40 mm × 40 mm were fabricated for this study. Mortars cured for 28 days were exposed to 0·2465 mol l-1 Na2 SO4 solution using accelerated ion migration test method for 36-h session using a 12V DC power source. Sulphate ion concentration was then determined through the ingressed mortar at 10 mm interval. A minimum water sorption gain of 0·61% was observed on the prism prepared with and cured in microbial solution. A maximum of 0·0289 and a minimum of 0·0093 water sorptivity coefficients were exhibited by the control prism and microbial prisms, respectively. The microbial prisms exhibited the lowest apparent diffusion coefficient (Dapp ) of 4·5179 × 10-11  m2  s-1 . CONCLUSIONS: Direct incorporation of B. megaterium in mortar preparation, curing or both regimes significantly retarded water sorption and lowered sulphate ion ingress. The inclusion of this bacterial in the mortar further complements the pozzolana pore structure benefits. SIGNIFICANCE AND IMPACT OF THE STUDY: This novel B. megaterium bacteria which can survive and cause biocementation within hydrating cement mortar when not encapsulated would result in a green innovation. Once adopted and applied in real-life scenario, it would promote construction of durable, safe, resilient and affordable shelter.

Antifungal Activity of Polyoxometalate-Ionic Liquids on Historical Brick.

Moulds inhabiting mineral-based materials may cause their biodeterioration, contributing to inestimable losses, especially in the case of cultural heritage objects and architectures. Fungi in mouldy buildings may also pose a threat to human health and constitute the main etiological factor in building related illnesses. In this context, research into novel compounds with antifungal activity is of high importance. The aim of this study was to evaluate the antifungal activity of polyoxometalate-ionic liquids (POM-ILs) and their use in the eradication of moulds from historical brick. In the disc diffusion assay, all the tested POM-ILs inhibited growth of a mixed culture of moulds including Engyodontium album, Cladosporium cladosporioides, Alternaria alternata and Aspergillus fumigatus. These were isolated from the surfaces of historical brick barracks at the Auschwitz II-Birkenau State Museum in Oswiecim, Poland. POM-IL coatings on historical brick samples, under model conditions, showed that two compounds demonstrated very high antifungal activity, completely limiting mould growth and development. The antifungal activity of the POM-ILs appeared to stem from their toxic effects on conidia, as evidenced by environmental scanning transmission electron microscopy observations. The results herein indicated that POM-ILs are promising disinfectant materials for use not only on historical objects, but probably also on other mineral-based materials.

Bacteria incorporated with calcium lactate pentahydrate to improve the mortar properties and self-healing occurrence.

Concrete can be harmful to the environment due to its high energy consumption and CO2 emission and also has a potential crack formation, which can promote a drop in its strength. Therefore, concrete is considered as a non-sustainable material. The mechanisms by which bacterial oxidation of organic carbon can precipitate calcite that may fill the voids and cracks on cement-based materials have been extensively investigated to prevent and heal the micro-cracks formation. Hence, this study focused on utilizing a new alkaliphilic bacterial strain indigenous to an Indonesian site, Lysinibacillus sphaericus strain SKC/VA-1, incorporated with calcium lactate pentahydrate, as a low-cost calcium source, with various bacterial inoculum concentrations. The bacterium was employed in this study due to its ability to adapt to basic pH, thus improving the physical properties and rejuvenating the micro-cracks. Experimentally, the addition of calcium lactate pentahydrate slightly affected the mortar properties. Likewise, bacteria-incorporated mortar exhibited an enhancement in the physical properties of mortar. The highest improvement of mechanical properties (an increase of 45% and 36% for compressive and indirect tensile strength, respectively) was achieved by the addition of calcium lactate pentahydrate incorporated with 10% v/v bacterial inoculum [about 7 × 107 CFU/ml (colony-forming unit/ml)]. The self-healing took place more rapidly on bacterial mortar supplemented with calcium lactate pentahydrate than on the control specimen. XRD analysis demonstrated that the mineralogical composition of self-healing precipitates was primarily dominated by calcite (CaCO3), indicating the capacity of L. sphaericus strain SKC/VA-1 to precipitate calcite through organic carbon oxidation for self-healing the artificial crack on the mortar. To our knowledge, this is the first report on the potential utilization of the bacterium L. sphaericus incorporated with calcium lactate pentahydrate to increase the mortar properties, including its self-healing ability. However, further study with the water-cement ratio variation is required to investigate the possibility of using L. sphaericus and calcium lactate pentahydrate as an alternative method rather than reducing the water-cement ratio to enhance the mortar properties.